Efficacy of green infrastructure in reducing exposure to local, traffic-related sources of airborne particulate matter (PM).

One aim of roadside green infrastructure (GI) is to mitigate exposure to local, traffic-generated pollutants. Here, we determine the efficacy of roadside GI in improving local air quality through the deposition and/or dispersion of airborne particulate matter (PM). PM was collected on both pumped air filters and on the leaves of a recently installed 'tredge' (trees managed as a head-high hedge) at an open road environment next to a primary school in Manchester, U.K. The magnetic properties of PM deposited on leaves and filters (size fractions PM10 and PM2.5) were deduced from hysteresis loops, first-order reversal curves (FORCs), and low-temperature remanence measurements. These were complemented with electron microscopy to identify changes in magnetic PM concentration downwind of the tredge/GI. We show that the tredge is permeable to airflow using a simple CO2 tracer experiment; hence, it allows interception and subsequent deposition of PM on its leaves. Magnetic loadings per m3 of air from filters (PM10 saturation magnetisation, Ms, at 5 K) were reduced by 40 % behind the tredge and a further 63 % in the playground; a total reduction of 78 % compared to roadside air. For the PM2.5 fraction, the reduction in magnetic loading behind the tredge was remarkable (82 %), reflecting efficient diffusional capture of sub-5 nm Fe-oxide particles by the tredge. Some direct mixing of roadside and playground air occurs at the back of the playground, caused by air flow over, and/or through gaps in, the slowly-permeable tredge. The magnetic loading on tredge leaves increased over successive days, capturing ~23 % of local, traffic-derived PM10. Using a heuristic two-dimensional turbulent mixing model, we assess the limited dispersion of PM < 22.5 µm induced by eddies in the tredge wake. This study demonstrates that PM deposition on leaves reduces exposure significantly in this school playground setting; hence, providing a cost-effective mitigation strategy.

Magnetic and microscopic investigation of airborne iron oxide nanoparticles in the London Underground.

Particulate matter (PM) concentration levels in the London Underground (LU) are higher than London background levels and beyond World Health Organization (WHO) defined limits. Wheel, track, and brake abrasion are the primary sources of particulate matter, producing predominantly Fe-rich particles that make the LU microenvironment particularly well suited to study using environmental magnetism. Here we combine magnetic properties, high-resolution electron microscopy, and electron tomography to characterize the structure, chemistry, and morphometric properties of LU particles in three dimensions with nanoscale resolution. Our findings show that LU PM is dominated by 5-500 nm particles of maghemite, occurring as 0.1-2 µm aggregated clusters, skewing the size-fractioned concentration of PM artificially to larger sizes when measured with traditional monitors. Magnetic properties are largely independent of the PM filter size (PM10, PM4, and PM2.5), and demonstrate the presence of superparamagnetic (< 30 nm), single-domain (30-70 nm), and vortex/pseudo-single domain (70-700 nm) signals only (i.e., no multi-domain particles > 1 µm). The oxidized nature of the particles suggests that PM exposure in the LU is dominated by resuspension of aged dust particles relative to freshly abraded, metallic particles from the wheel/track/brake system, suggesting that periodic removal of accumulated dust from underground tunnels might provide a cost-effective strategy for reducing exposure. The abundance of ultrafine particles identified here could have particularly adverse health impacts as their smaller size makes it possible to pass from lungs to the blood stream. Magnetic methods are shown to provide an accurate assessment of ultrafine PM characteristics, providing a robust route to monitoring, and potentially mitigating this hazard.

Cooperativity between Sertoli cells and testicular peritubular cells in the production and deposition of extracellular matrix components.

We examined the synthesis and deposition of extracellular matrix (ECM) components in cultures of Sertoli cells and testicular peritubular cells maintained alone or in contact with each other. Levels of soluble ECM components produced by populations of isolated Sertoli cells and testicular peritubular cells were determined quantitatively by competitive enzyme-linked immunoabsorbent assays, using antibodies shown to react specifically with Type I collagen, Type IV collagen, laminin, or fibronectin. Peritubular cells in monoculture released into the medium fibronectin (432 to 560 ng/microgram cell DNA per 48 h), Type I collagen (223 to 276 ng/microgram cell DNA per 48 h), and Type IV collagen (350 to 436 ng/microgram cell DNA per 48 h) during the initial six days of culture in serum-free medium. In contrast, Sertoli cells in monoculture released into the medium Type IV collagen (322 to 419 ng/microgram cell DNA per 48 h) but did not form detectable amounts of Type I collagen or fibronectin during the initial six days of culture. Neither cell type produced detectable quantities of soluble laminin. Immunocytochemical localization investigations demonstrated that peritubular cells in monoculture were positive for fibronectin, Type I collagen, and Type IV collagen but negative for laminin. In all monocultures most of the ECM components were intracellular, with scant deposition as extracellular fibrils. Sertoli cells were positive immunocytochemically for Type IV collagen and laminin but negative for fibronectin and Type I collagen. Co-cultures of peritubular cells and Sertoli cells resulted in interactions that quantitatively altered levels of soluble ECM components present in the medium. This was correlated with an increased deposition of ECM components in extracellular fibrils. The data correlated with an increased deposition of ECM components in extracellular fibrils. The data presented here we interpret to indicate that the two cell types in co-culture act cooperatively in the formation and deposition of ECM components. Results are discussed with respect to the nature of interactions between mesenchymal peritubular cell precursors and adjacent epithelial Sertoli cell precursors in the formation of the basal lamina of the seminiferous tubule.

CaLB: a 43 amino acid calcium-dependent membrane/phospholipid binding domain in p120 Ras GTPase-activating protein.

CaLB was originally observed as a conserved sequence motif in various calcium-responsive signalling proteins and also in p120 Ras GTPase activating protein (p120GAP) (Clark et al. Cell 65: 1043-1051, 1991). Here we show the 43 residue CaLB motif in p120GAP is a functional protein domain that when expressed as a fusion protein in vitro confers Ca(2+)-dependent interactions with cellular membranes and phosphatidylserine and phosphatidylinositol vesicles. p120GAP, but not a mutant lacking the CaLB domain, associates with the particulate fraction of cells in response to elevated intracellular Ca2+ suggesting that p120GAP may be regulated in part by calcium signals. Addition of the p120GAP CaLB domain was able to restore transforming activity and particulate localization to an otherwise transformation-defective and cytosolic mutant v-Sre tyrosine kinase. The CaLB domain appears to be a prevalent protein module that may affect the molecular interactions and subcellular localization of signalling proteins.

Substrate and cofactor binding to fluorescently labeled cytoplasmic malate dehydrogenase.

Cytoplasmic malate dehydrogenase (cMDH) is a key enzyme in several metabolic pathways. Though its activity has been examined extensively, there are lingering mechanistic uncertainties involving substrate and cofactor binding. To more completely understand this enzyme's interactions with cofactor and substrate ligands, a fluorescent reporter group was introduced into the enzyme's structure. This was accomplished by selective modification of Cys 110. The reaction placed an aminonaphthaline sulfonic acid group near the enzyme's active site. Substrate, inhibitor, and NAD binding activities were characterized using changes in this label's fluorescence. Results demonstrated that both substrate and cofactor molecules bound to the enzyme in the absence of their companion ligands. This is in contrast to strictly ordered cofactor then substrate binding as has been suggested by kinetic analyses of closely related enzymes. Binding results also indicated that the cofactor, NAD, bound to cMDH in a negatively cooperative manner, but substrates and the inhibitor, hydroxymalonate, bound non-cooperatively. Multiple substrate binding modes were identified and interactions between substrate and cofactor binding were found.

Importance of the Chiral Centers of Jasmonic Acid in the Responses of Plants (Activities and Antagonism between Natural and Synthetic Analogs).

The importance of the two chiral centers at C-3 and C-7 in the molecular structure of jasmonic acid in plant responses was investigated. We separated methyl jasmonate (MeJA) into (3R)- and (3S)-isomers with a fixed stereochemistry at C-3, but epimerization at C-7 is possible. The four isomers of the nonepimerizable analog 7-methyl MeJA were synthesized. These six esters and their corresponding acids were tested in three bioassays: (a) senescence in sunflower (Helianthus annuus) cotyledons; (b) proteinase inhibitor II gene expression in transgenic tobacco (Nicotiana tabacum) with [beta]-glucuronidase as a biochemical reporter; and (c) seed germination in Brassica napus and wheat (Triticum aestivum). The esters and acids had similar activities in the three assays, with the ester being more effective than its acid. The (3R)-stereochemistry was critical for jasmonate activity. Although activity was reduced after substituting the C-7 proton with a methyl group, the analogs with (3R,7R)- or (3R,7S)-stereochemistry were active in some of the assays. Although the four isomers of 7-methyl MeJA were inactive or only weakly active in the senescence assay, they could overcome the senescence-promoting effect of (3R)-MeJA. The strongest antagonistic effect was observed with the (3R,7S)-isomer.

Effects of weight bearing and non-weight bearing exercises on bone properties using calcaneal quantitative ultrasound.

OBJECTIVE: This study was designed to investigate bone properties using heel quantitative ultrasound (QUS) in young adults participating in various sports. METHODS: A cross sectional study was performed on Chinese male students (n = 55), aged 18-22 years. Subjects with previous fractures or suffering from any diseases known to affect bone metabolism or taking any medication with such an effect, were not included. The subjects were categorised according to their main sporting activities, including soccer (n = 15) (a high impact, weight bearing exercise), dancing (n = 10) (a low impact, weight bearing exercise), and swimming (n = 15) (non-weight bearing exercise). A sedentary group acted as controls (n = 15). A reproducibility study of the velocity of sound (VOS) and the broadband ultrasound attenuation (BUA) measurement was performed and analysed using the intraclass correlation coefficient (ICC). RESULTS: There was good intra-investigator and inter-investigator agreement (ICC > or = 0.8; p < 0.05) in the measurement of BUA and VOS. No significant differences in BUA and VOS (p > 0.05) were found between the dominant and non-dominant heel. Soccer players (137 +/- 4.3 dB/MHz; 1575 +/- 56 m/s; 544.1 +/- 48.4) and dancers (134.6 +/- 3.7 dB/MHz; 1538 +/- 46 m/s; 503.0 +/- 37.0) had significantly higher BUA, VOS, and stiffness index (SI) scores (p < 0.05), respectively, than swimmers (124.1 +/- 5.1 dB/MHz; 1495 +/- 42 m/s; 423.3 +/- 46.9) and the sedentary control group (119.9 +/- 6.1 dB/MHz; 1452 +/- 41 m/s; 369.9 +/- 46.4). A trend of a significant linear increase with the weight bearing and high impact exercise was revealed in all QUS parameters (p < 0.05). CONCLUSION: This cross sectional study indicated that regular participation in weight bearing exercise in young people might be beneficial for accruing peak bone mass and optimising bone structure.

Epigenetic mechanisms of tumorigenicity manifesting in stem cells.

One of the biggest roadblocks to using stem cells as the basis for regenerative medicine therapies is the tumorigenicity of stem cells. Unfortunately, the unique abilities of stem cells to self-renew and differentiate into a variety of cell types are also mechanistically linked to their tumorigenic behaviors. Understanding the mechanisms underlying the close relationship between stem cells and cancer cells has therefore become a primary goal in the field. In addition, knowledge gained from investigating the striking parallels between mechanisms orchestrating normal embryogenesis and those that invoke tumorigenesis may well serve as the foundation for developing novel cancer treatments. Emerging discoveries have demonstrated that epigenetic regulatory machinery has important roles in normal stem cell functions, cancer development and cancer stem cell (CSC) identity. These studies provide valuable insights into both the shared and distinct mechanisms by which pluripotency and oncogenicity are established and regulated. In this review, the cancer-related epigenetic mechanisms found in pluripotent stem cells and cancer cells will be discussed, focusing on both the similarities and the differences.

Modulation of Sertoli cell functions in the two-chamber assembly by peritubular cells and extracellular matrix.

A two-chamber assembly has been employed to investigate influences of peritubular cells (PC) and extracellular matrix (Matrigel) on barrier formation by Sertoli cells (SC) in culture and on transferrin production. The kinetics of transport of [3H]inulin across a Millipore filter were essentially the same in the presence or absence of Matrigel or PC. In contrast, a SC monolayer retarded the diffusion of [3H]inulin, increasing the estimated time for 50% equilibration from about 4 h to approximately 12 h. Matrigel and PC each independently further increased the equilibration time, with the largest effects elicited by the presence of Matrigel (approximately 21 h). Data have been interpreted to indicate that these two components, especially extracellular matrix, facilitate the formation of a functional barrier by SC in the two-chamber system. PC assume a more important role than Matrigel in the modulation of transferrin secretion by SC. Transferrin concentrations were higher in the inner chamber, corresponding to those in the adluminal compartment, but transferrin masses were higher in the outer chamber under the conditions described. We report the effects of the presence and absence of Matrigel, PC, and FSH on levels of transferrin secreted by SC. Addition of FSH resulted in increased transferrin secretion by SC maintained under all conditions examined. We compare our data with those previously reported by others and attempt to provide a basis for the differences observed. We discuss the properties of the system and outline major advantages and limits of the two-chamber assembly in investigations on the polarity and properties of SC in culture.

Nephropathy in non-insulin-dependent diabetes mellitus.

End-stage renal disease develops in about 5 percent of patients with non-insulin-dependent diabetes mellitus (NIDDM). The large majority of diabetic patients have this form of the disease. Thus, end-stage renal disease is an important clinical problem in patients with NIDDM. Moreover, hypertension and its macrovascular sequelae are significant problems in patients with NIDDM and may be linked with renal disease. A review of the problem of nephropathy in NIDDM is attempted, pointing out, where data are available, the clinical and pathophysiologic differences from its presentation in insulin-dependent diabetes. The need for further studies of the impact of renal disease in this maturity onset form of diabetes is emphasized.

Specific immunohistochemical localization of osteonectin and collagen types I and III in fetal and adult porcine dental tissues.

Affinity-purified antibodies have been used in combination with the peroxidase-antiperoxidase technique to study the distribution of osteonectin and collagen types I and III in porcine dental tissues. Tissue sections (2 mm thick), including unerupted (fetal) or erupted (adult) teeth, were fixed in periodate-lysine-paraformaldehyde, demineralized in 12% w/v ethylenediaminetetraacetic acid, and after embedding, 6 micron sections were prepared for immunolocalization. Strong staining for osteonectin was observed in dentine of unerupted teeth and in the associated alveolar bone. Light to moderate staining was observed in the dental pulp, stratum intermedium, stellate reticulum, and the reticular elements in the endosteal spaces. In erupted teeth, osteonectin staining in dentine was concentrated around dentinal tubules and the associated alveolar bone stained with variable intensity. Cementum was poorly stained. However, the periodontal ligament and reticular material in the endosteal spaces showed moderate to strong staining. Weaker staining was apparent in the pulp and lamina propria of the gingiva. In comparison, type I collagen showed a similar distribution to osteonectin in both fetal and adult tissues, whereas type III collagen was generally restricted to the periodontal ligament, reticular elements of the endosteal spaces, and Sharpey's fibers in bone and cementum. Both odontoblast and ameloblast layers in fetal tissues stained for osteonectin and type III collagen.

An objective sperm cytotoxicity assay for male-specific antisera based on ATP levels of unlysed cells: application to assay of H-Y antigen.

We correlated the decrease in levels of ATP in spermatozoa with the extent of cytotoxicity elicited by antibodies against antigenic components on sperm. In the presence of concentrations of complement which did not cause cytolysis or influence the ATP content of epididymal sperm, addition of heat-in-activated sera from non-immunized mice, rats or rabbits did not result in sperm cytolysis or a fall in ATP content. In contrast, addition of rabbit anti-rat spermatocyte sera, which has previously been shown to react with rat spermatozoa (Tung, P.S. and Fritz, I.B. (1978) Dev. Biol. 64, 297-315), did cause sperm cytolysis and a decrease in ATP content. The titre of this antiserum for 50% cytolysis was between 1 : 128 and 1 : 256, as determined by the fall in ATP content. Using these criteria, we examined the cytotoxicity against sperm of different samples of anti H-Y sera. We examined the influence of monoclonal antibody against H-Y, mouse H-Y antisera and rat H-Y antisera raised in inbred females immunized with spleen cells from males of the same strains. In all cases, anti-H-Y lowered ATP levels and lysed sperm with a cytotoxic titre between 1 : 8 and 1 : 16. Measurements of the decrease in ATP content in sperm have been shown to provide an objective and reliable estimate of the percentage of spermatozoa lysed by H-Y antisera. Cytotoxic activity of H-Y antisera was removed by absorption with spleen cells from male mice but not by absorption with spleen cells from female mice.

Pancreatic insulinomas. A 15-year experience.

OBJECTIVE: To describe our experience in the management of patients with pancreatic insulinomas, emphasizing the need for preoperative localization and the outcome of surgical treatment. DESIGN: A case series. SETTING: A university hospital in Hong Kong. PATIENTS: From 1981 to 1995, 27 patients with pancreatic insulinomas were surgically treated; the mean follow-up was 25.4 months. MAIN OUTCOME MEASURES: Postoperative morbidity and euglycemia during the follow-up period. RESULTS: The accuracy of tumor localization by ultrasonography, computed tomography, and angiography was 33%, 44%, and 52%, respectively. Venous sampling for an insulin assay regionalized 90% of the tumors. In 24 patients with solitary tumors, most lesions detected or missed by preoperative localization could be either seen (n = 14) or palpated (n = 22). Intraoperative ultrasonography (n = 17) has been routinely performed since 1987; nonpalpable tumors were imaged in 2 of 15 patients with solitary tumors. Eight solitary occult tumors were detected by palpation alone or a combination of palpation and intraoperative ultrasonography. Operative mortality occurred in 1 (3.7%) of the 27 patients, while major morbidity developed in 9 (33%) of the patients. Euglycemia was achieved in 25 patients. Surgery cured all patients with benign insulinomas, whereas the cure rate for patients with malignant neoplasms was only 33%. The type of surgical treatment or correct preoperative localization did not affect the outcome of surgery. CONCLUSIONS: Pancreatic insulinomas can be readily localized intraoperatively despite failed preoperative localization studies. Surgical treatment cured benign adenomas but was associated with notable morbidity.

Thoracoscopic esophageal mobilization for pharyngolaryngoesophagectomy.

BACKGROUND: Pharyngolaryngoesophagectomy (PLE) for hypopharyngeal cancers and tumors of the cervical esophagus is a procedure of significant morbidity and mortality. Conventional esophageal mobilization is performed with the transhiatal dissection technique. Thoracoscopic esophageal mobilization is tested as an alternative to determine whether surgical outcome can be improved. METHODS: From 1994 to 1998, thoracoscopic mobilization was carried out in 30 consecutive patients who underwent PLE (PLE-TS). This was compared to a historical cohort of 30 patients who had PLE with transhiatal mobilization (PLE-TH). RESULTS: In PLE-TS, thoracoscopic esophageal mobilization was successful in 28 patients (93%). Median blood loss was 700 mL (range, 164 to 3,000 mL) compared to 1,000 mL (range, 400 to 2,200 mL) in group PLE-TH, p = 0.21. Thoracoscopy time was 90 minutes (range, 60 to 180 minutes). Total operating time were 392 minutes (range, 180 to 570 minutes) and 300 minutes (range, 150 to 550 minutes) in PLE-TS and PLE-TH, respectively (p = 0.03). Major pulmonary complications occurred in 7 (23%) and 8 (27%) patients in PLE-TS and PLE-TH, respectively (p = 0.77). Cardiac complications occurred in 7 (23%) and 5 (17%) patients in PLE-TS and PLE-TH, respectively (p = 0.52). Thirty-day mortality rates were 3.3% and 10% (p = 0.6) and hospital mortality rates were 13% and 17%, (p = 1.0). CONCLUSIONS: Thoracoscopy was safe and feasible. Morbidity and mortality after PLE was not significantly reduced. The theoretical advantage offered by thoracoscopy may be offset by the lengthened time of one-lung anesthesia.

Characterization and use of a recombinant retroviral system for the analysis of drug resistant HIV.

A recombinant retroviral system was used for the analysis of early HIV breakthrough infection in the presence of antiviral drugs. The use of replication-defective HIV allowed a quantitative analysis of a single cycle of infection. This report characterizes this recombinant HIV system and demonstrates it's validity in comparison to standard assays. It is demonstrated that the protease inhibitor XM323 inhibits both early and late events in the HIV life-cycle, while dextran sulphate inhibits only early events. In addition, it is shown that this system can be used for detecting and quantitating drug resistant HIV. Thus, the use of this system may provide both novel information about the stage of the viral life-cycle inhibited and a preliminary assessment of the mechanism(s) responsible for breakthrough infection in the presence of antiretroviral drugs.

Laparoscopic transperitoneal clipping of the inferior mesenteric artery for the management of type II endoleak after endovascular repair of an aneurysm.

We report the case of a high risk patient with an abdominal infrarenal aortic aneurysm (AAA) who was treated by endovascular technique and the subsequent management of a type II endoleak by the laparoscopic approach. In this case, a 74-year-old woman with a 6-cm infrarenal AAA underwent endovascular repair using a bifurcated stent-graft device. Surveillance CT scan showed a persistent type II endoleak at 1 week and 3 months after the operation. Angiography confirmed retrograde flow from the inferior mesenteric artery (IMA). Attempted transarterial embolization of the IMA via the superior mesenteric artery was not successful. Laparoscopic transperitoneal IMA clipping was performed. Subsequent aortic duplex scan and CT scan confirmed complete elimination of the type II endoleak. We conclude that a combination of endovascular and laparoscopic procedures can be used to manage AAA successfully.

Esophageal small cell carcinomas: clinicopathologic parameters, p53 overexpression, proliferation marker, and their impact on pathogenesis.

OBJECTIVE: To evaluate the clinicopathologic features and the roles of p53 and MIB-1 in esophageal small cell carcinoma. METHOD: Twenty patients (14 men and 6 women) with esophageal small cell carcinoma treated in our hospital from 1982 through 1996 were studied. The clinicopathologic features, treatment received, and survival data of these patients were documented. Representative tissue was collected from each tumor, and immunohistochemical preparations for p53 protein and MIB-1 were made. RESULTS: Small cell carcinoma accounted for 1.3% of all esophageal malignant tumors. The median age of patients at presentation was 60 years. On gross examination, the tumors were large ulcerative lesions (median length, 7.5 cm). In 17 patients in whom p53 immunohistochemical study was performed, p53 protein was detected in 65% (9 of 17). All stage IV tumors were negative for p53 expression. The median tumor cell MIB-1 score was high at 855 (range, 810-964) positive cells per 1000. Overall median survival was 3.4 months. In patients who underwent chemotherapy, there was significant response. CONCLUSIONS: Esophageal small cell carcinoma is an aggressive tumor. Overexpression of p53 is associated with early stages of carcinogenesis. The high proliferative index, as defined by the MIB-1 immunohistochemical method, may be related to aggressive behavior and high sensitivity to chemotherapy and radiotherapy.

Gastric rupture after Heimlich maneuver and cardiopulmonary resuscitation.

Choking is a common emergency problem. The Heimlich maneuver is unquestionably effective in relieving airway obstruction. Serious and life-threatening complications may arise, however, if the maneuver is applied incorrectly. Two cases of gastric rupture after Heimlich maneuver are reported. Lay public, paramedics and the medical professionals should be educated with the correct technique of Heimlich maneuver and its potential complications. All patients receiving Heimlich maneuver should be examined by an experienced physician.